Dendrimer modified biochip for detection of 2,4,6 trinitrotoluene on SPR immunosensor: Fabrication and advantages

Praveen Singh, Takeshi Onodera, Yutaka Mizuta, Kiyoshi Matsumoto, Norio Miura, Kiyoshi Toko

研究成果: ジャーナルへの寄稿学術誌査読

63 被引用数 (Scopus)

抄録

This paper describes the fabrication and evaluation of new sensor chip technology based on application of polyamidoamine (PAMAM) dendrimer generation 4 (G4) for enhancing surface loading capacity and efficiency for selective and sensitive detection of explosive molecule trinitrotoluene (TNT) under competitive inhibition assay format. The fabrication process of a nano-scale biosensor chip surface was optimized at various steps involving thiol layer, cross-linker, dinitro phenylated-keyhole limpet hemocyanin (DNP-KLH) protein conjugate as ligand. The top ligand layer is supported by underlying 11-amino 1-undecanethiol hydrochloride (AUT) self-assembled monolayer (SAM) and PAMAM dendrimer G4. Bis-sulfosuccinimidyl suberate (BS3) was used as homobifunctional crosslinker to react with amine groups at both the ends. PAMAM dendrimer activation was achieved over 11-amino 1-undecanethiol SAMs over gold chip. PAMAM sensor surfaces scored many advantages over the planer surfaces such as enhanced antibody immobilization, allowing the use of lower concentrated antibody to work under the regime of lower detection limit besides stable and robust surface leading to longer life. Bioactive thin films were fabricated over gold chip via layer-by-layer self-assembly methods. Biomolecular interaction between DNP-KLH conjugate surface and specific TNP-gly-KLH mouse IgG antibody was monitored through surface plasmon resonance (SPR) based real-time optical transducer. The quantitation of TNT on this bioactive surface was done using the competitive inhibition immunoassay. The DNP-KLH surface biosensor has shown a detection limit of 110 ppt for TNT molecule. This TNT specific biosensor holds the promise to be most sensitive, easy to regenerate, highly stable, low response loss sensor surface under competitive assay format. A 12 s injection of 10 mM glycine-HCl solution or 50 mM NaOH solution was found adequate for regeneration of DNP-KLH surface for repeated use. DNP-KLH sensor platform was checked for its repeatability and storability.

本文言語英語
ページ(範囲)403-409
ページ数7
ジャーナルSensors and Actuators, B: Chemical
137
2
DOI
出版ステータス出版済み - 4月 2 2009

!!!All Science Journal Classification (ASJC) codes

  • 電子材料、光学材料、および磁性材料
  • 器械工学
  • 凝縮系物理学
  • 表面、皮膜および薄膜
  • 金属および合金
  • 電子工学および電気工学
  • 材料化学

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